The proposed work is designed to study the reversible interaction of alcohol dehydrogenase with F-actin, and the effect of relevant metabolites on their interaction. We will test whether the catalytic properties of the bound enzyme differ from those of the free enzyme, and whether the free and bound enzyme have different proteolytic susceptibility. Both the horse liver and yeast isozymes will be used. The interaction will be studied by separating the free and bound enzyme using Ultracentrifugation and assaying the activity of the free enzyme in the supernatant. The effect of F-actin on the turnover rate of alcohol dehydrogenase will be studied by comparing the rate of trypsin and chymotrypsin digestion of the enzyme in the presence and absence of F-actin. We are proposing to study alcohol dehydrogenase in a system more similar to a cellular environment than a buffer solution. Our work will provide a more realistic understanding of the enzymatic properties of alcohol dehydrogenase and the oxidation of alcohol in liver and yeast cells in vivo. Our long term objective is to study the effect of changing the environment and the cytoskeletal components on the biological activity of cytoplasmic enzymes.